In vitro placenta barrier model using primary human trophoblasts, underlying connective tissue and vascular endothelium.

Fetal development may be compromised by adverse events at the placental interface between mother and fetus. However, it is still unclear how the communication between mother and fetus occurs through the placenta. In vitro - models of the human placental barrier, which could help our understanding and which recreate three-dimensional (3D) structures with biological functionalities and vasculatures, have not been reported yet. Here we present a 3D-vascularized human primary placental barrier model which can be constructed in 1 day. We illustrate the similarity of our model to first trimester human placenta, both in its structure and in its ability to respond to altered oxygen and to secrete factors that cause damage cells across the barrier including embryonic cortical neurons. We use this model to highlight the possibility that both the trophoblast and the endothelium within the placenta might play a role in the fetomaternal dialogue.

Nanoparticle-induced neuronal toxicity across placental barriers is mediated by autophagy and dependent on astrocytes.

The potential for maternal nanoparticle (NP) exposures to cause developmental toxicity in the fetus without the direct passage of NPs has previously been shown, but the mechanism remained elusive. We now demonstrate that exposure of cobalt and chromium NPs to BeWo cell barriers, an in vitro model of the human placenta, triggers impairment of the autophagic flux and release of interleukin-6. This contributes to the altered differentiation of human neural progenitor cells and DNA damage in the derived neurons and astrocytes. Crucially, neuronal DNA damage is mediated by astrocytes. Inhibiting the autophagic degradation in the BeWo barrier by overexpression of the dominant-negative human ATG4BC74A significantly reduces the levels of DNA damage in astrocytes. In vivo, indirect NP toxicity in mice results in neurodevelopmental abnormalities with reactive astrogliosis and increased DNA damage in the fetal hippocampus. Our results demonstrate the potential importance of autophagy to elicit NP toxicity and the risk of indirect developmental neurotoxicity after maternal NP exposure.

Treating the placenta to prevent adverse effects of gestational hypoxia on fetal brain development.

Some neuropsychiatric disease, including schizophrenia, may originate during prenatal development, following periods of gestational hypoxia and placental oxidative stress. Here we investigated if gestational hypoxia promotes damaging secretions from the placenta that affect fetal development and whether a mitochondria-targeted antioxidant MitoQ might prevent this. Gestational hypoxia caused low birth-weight and changes in young adult offspring brain, mimicking those in human neuropsychiatric disease. Exposure of cultured neurons to fetal plasma or to secretions from the placenta or from model trophoblast barriers that had been exposed to altered oxygenation caused similar morphological changes. The secretions and plasma contained altered microRNAs whose targets were linked with changes in gene expression in the fetal brain and with human schizophrenia loci. Molecular and morphological changes in vivo and in vitro were prevented by a single dose of MitoQ bound to nanoparticles, which were shown to localise and prevent oxidative stress in the placenta but not in the fetus. We suggest the possibility of developing preventative treatments that target the placenta and not the fetus to reduce risk of psychiatric disease in later life.

Secretions from placenta, after hypoxia/reoxygenation, can damage developing neurones of brain under experimental conditions.

Some psychiatric diseases in children and young adults are thought to originate from adverse exposures during foetal life, including hypoxia and hypoxia/reoxygenation. The mechanism is not understood. Several authors have emphasised that the placenta is likely to play an important role as the key interface between mother and foetus. Here we have explored whether a first trimester human placenta or model barrier of primary human cytotrophoblasts might secrete factors, in response to hypoxia or hypoxia/reoxygenation, that could damage neurones. We find that the secretions in conditioned media caused an increase of [Ca(2+)]i and mitochondrial free radicals and a decrease of dendritic lengths, branching complexity, spine density and synaptic activity in dissociated neurones from embryonic rat cerebral cortex. There was altered staining of glutamate and GABA receptors. We identify glutamate as an active factor within the conditioned media and demonstrate a specific release of glutamate from the placenta/cytotrophoblast barriers invitro after hypoxia or hypoxia/reoxygenation. Injection of conditioned media into developing brains of P4 rats reduced the numerical density of parvalbumin-containing neurones in cortex, hippocampus and reticular nucleus, reduced immunostaining of glutamate receptors and altered cellular turnover. These results show that the placenta is able to release factors, in response to altered oxygen, that can damage developing neurones under experimental conditions.

RSL Fusion with Excision of Distal Scaphoid and Triquetrum: A Cadaveric Study.

Radioscapholunate (RSL) fusion has been utilized for treatment of radiocarpal arthritis for patients with an intact midcarpal joint. This preserves midcarpal joint motion while alleviating pain. Dart thrower's motion (DTM), which has been emphasized recently, is mainly a midcarpal joint motion. Question A cadaveric study was designed to measure and compare the range of motion (ROM) of the human wrist before and after an RSL fusion, followed by distal scaphoid excision, and finally excision of the triquetrum. Methods Twelve embalmed adult cadaveric upper limbs were assessed. The wrist motion was measured with an electrogoniometer. Measurements of the flexion-extension plane and radial-ulnar deviation plane were obtained for baseline after capsulotomy, after simulated RSL fusion with memory staples, after distal scaphoidectomy, and after excision of the triquetrum. Results The effects of scaphoid and triquetrum excision were expressed as improvements in movement over that of the preceding step. RSL fusion alone resulted in a decrease of the flexion-extension (F-E) arc by 36% and the radioulnar deviation (R-U) arc by 30%. Excision of the distal scaphoid with RSL improved the F-E arc by 34% and the R-U arc by 34%. With excision of the triquetrum, the F-E arc improved further by 13% and the R-U arc by 21%. The ROM of the simulated RSL fusion with distal scaphoidectomy is improved with excising the triquetrum, mainly through an increase in ulnar deviation motion. RSL fusion with distal scaphoidectomy and triquetrectomy can be an alternative to total wrist arthrodesis for patients with an intact midcarpal joint.

Consequences of exposure to peri-articular injections of micro- and nano-particulate cobalt-chromium alloy.

Metal hip replacements generate both metal particles and ions. The biological effects of peri-articular exposure to nanometre and micron sized cobalt chrome (CoCr) wear particles were investigated in a mouse model. Mice received injections of two clinically relevant doses of nanoparticles (32 nm), one of micron sized (2.9 µm) CoCr particles or vehicle alone into the right knee joint at 0, 6, 12 and 18 weeks. Mice were analysed for genotoxic and immunological effects 1, 4 and 40 weeks post exposure. Nanoparticles but not micron particles progressively corroded at the injection site. Micron sized particles were physically removed. No increase of Co or Cr was seen in peripheral blood between 1 and 40 weeks post exposure to particles. No significant inflammatory changes were observed in the knee tissues including ALVAL or necrosis. DNA damage was increased in bone marrow at one and forty weeks and in cells isolated from frontal cortex at 40 weeks after injection with nanoparticles. Mice exposed to the micron sized, but not nanoparticles became immunologically sensitized to Cr(III), Cr (VI) and Ni(II) over the 40 week period as determined by lymphocyte transformation and ELISpot (IFN-γ and IL-2) assays. The data indicated that the response to the micron sized particles was Th1 driven, indicative of type IV hypersensitivity. This study adds to understanding of the potential adverse biological reactions to metal wear products.

Thresholds for indirect DNA damage across cellular barriers for orthopaedic biomaterials.

Cobalt-chromium particles and ions can induce indirect DNA damage and chromosome aberrations in human cells on the other side of a cellular barrier in tissue culture. This occurs by intercellular signalling across the barrier. We now show that the threshold for this effect depends on the metal form and the particle composition. Ionic cobalt and chromium induced single strand breaks at concentrations equivalent to those found in the blood of patients with well functioning metal on metal hip prostheses. However, they only caused double strand breaks if the chromium was present as chromium (VI), and did not induce chromosome aberrations. Nanoparticles of cobalt-chromium alloy caused DNA double strand breaks and chromosome aberrations, of which the majority were tetraploidy. Ceramic nanoparticles induced only single strand breaks and/or alkaline labile sites when indirectly exposed to human fibroblasts. The assessment of reproductive risk from maternal exposure to biomaterials is not yet possible with epidemiology. Whilst the barrier model used here differs from the in vivo situation in several respects, it may be useful as a framework to evaluate biomaterial induced damage across physiological barriers.

Nanoparticles can cause DNA damage across a cellular barrier.

The increasing use of nanoparticles in medicine has raised concerns over their ability to gain access to privileged sites in the body. Here, we show that cobalt-chromium nanoparticles (29.5 +/- 6.3 nm in diameter) can damage human fibroblast cells across an intact cellular barrier without having to cross the barrier. The damage is mediated by a novel mechanism involving transmission of purine nucleotides (such as ATP) and intercellular signalling within the barrier through connexin gap junctions or hemichannels and pannexin channels. The outcome, which includes DNA damage without significant cell death, is different from that observed in cells subjected to direct exposure to nanoparticles. Our results suggest the importance of indirect effects when evaluating the safety of nanoparticles. The potential damage to tissues located behind cellular barriers needs to be considered when using nanoparticles for targeting diseased states.

Translunate fracture with associated perilunate injury: 3 case reports with introduction of the translunate arc concept.

We report 3 cases of translunate fractures with associated perilunate dislocations (or subluxation). We believe the translunate injury reflects a higher-velocity trauma and produces further destabilization of the carpus when compared with the established greater and lesser arc injuries. A modification to Johnson's perilunate injury classification system is proposed: the addition of a translunate arc injury subgroup, which would include all perilunate injuries with translunate fractures.

Clinical results of coracoacromial ligament transfer in acromioclavicular dislocations: A review of published literature.

Acromioclavicular joint dislocations are common injuries, which typically occur with trauma in young men. Treatment recommendations for these injuries are highly variable and controversial. There are greater than 100 surgical techniques described for operative treatment of this injury. One of the most widely recommended methods of surgical reconstruction for acromioclavicular joint dislocations is to utilize the coracoacromial ligament for stabilization of the distal clavicle. Several modifications of this procedure have been described which have involved adjunct coracoclavicular fixation or fixation across acromioclavicular joint. Although the literature is replete with descriptive papers, there is paucity of studies evaluating the surgical outcome of this procedure. We systematically reviewed the English language published literature in peer reviewed journals (Medline, EMBASE, SCOPUS) and assigned a level of evidence for available studies. We critically reviewed each paper for the flaws and biases and then evaluated the comparable clinical outcomes for various procedures and their modifications. The published literature consists entirely of case series (Level IV evidence) with variability in surgical technique and outcome measures. On review there is low level evidence to support the use of coracoacromial ligament for acromioclavicular dislocation but it has been associated with high rate of deformity recurrence. Adjunct fixation does not improve clinical results when compared to isolated coracoacromial ligament transfer. This is in part because of the high incidence of fixation related complications. Similar results are reported with coracoacromial ligament reconstruction for acute and chronic cases. The development of secondary acromioclavicular joint symptoms with distal clavicle retention is poorly reported with the incidence rate varying from 12% to 32%. Despite this, the retention or excision of distal clavicle did not affect overall clinical results except in the patients with pre existing acromioclavicular joint osteoarthritis who have inferior results with retention of distal end of clavicle. Further well designed clinical trials with validated outcome measures are required to fully evaluate the clinical results of this procedure.

Genotoxic effects of particles of surgical cobalt chrome alloy on human cells of different age in vitro.

Humans are exposed to metals from industry, the environment and from wear debris from worn orthopaedic joint replacements. Patients exposed to worn cobalt chrome hip replacements show an increase of chromosome aberrations in the bone marrow adjacent to the implant and an increase of chromosome translocations and aneuploidy in the peripheral blood. This study has tested whether particles of surgical cobalt chrome alloy are able to induce similar DNA damage and chromosome aberrations in human cells in vitro. Because increasingly young patients are receiving hip replacements it has also tested whether the response is altered at different cellular age in vitro. Primary human fibroblasts, were tested at different pre senescent population doublings (PD10 (young) and PD35 (older)) to particles of cobalt chrome alloy for up to 15 days. As in patients there was an increase of aneuploidy, chromosome translocations and DNA damage after exposure to the cobalt chrome particles in vitro. The overall level of DNA damage and numerical and structural aberrations was approximately the same in young and older cells. However, the cellular reaction to the DNA damage was different. Older cells showed a greater loss of viability and induction of senescence and a lesser rate of mitosis and cell growth than young cells. They showed less change in transcription, particularly of p38 and caspase 10 mRNA levels, than young cells. They showed more complex aneuploidy in association with unseparated or prematurely separated chromatids. This study suggests that at least part of the chromosome changes in patients with worn implants may be due to direct effects of the metal wear particles from the implant. It would be of interest to test whether the altered reaction of the human cells at different in vitro age might correspond with a different incidence of chromosome aberrations in patients at different ages.

Psychiatric morbidity in non-psychiatric geriatric inpatients.

AIM: To evaluate the profile of psychiatric disorders in geriatric inpatients. METHODS: A total of 528 patients (age 65 years and above) admitted to various departments of the teaching hospital attached to the Government Medical College, Amritsar from 15 September 2001 to 14 September 2002 were included in the study. Psychiatric assessment of patients was made on the basis of psychogeriatric assessment scales (PAS) and present state examination (PSE-ninth edition, 1974). The ICD-10 criteria were used for psychiatric diagnoses. General medical conditions were diagnosed by consultants of the respective departments. The patients were finally assessed by the consultant of the Department of Psychiatry. The obtained data were analysed using the chi-square test. RESULTS: Of the 528 patients, 260 (49%) had psychiatric co-morbidity. The most common psychiatric disorder was depression (25.94%), followed by adjustment disorders (11%), anxiety disorders (4.54%), dementias (3.6%), delirium (3%), bipolar disorders (0.8%), and substance-related disorders (0.4%). CONCLUSION: The above findings emphasize the importance of consultation-liaison psychiatry, especially in geriatric patients.

A prospective longterm study of primary chronic angle closure glaucoma.

PURPOSE: To prospectively evaluate the longterm outcome of therapy for chronic primary angle closure glaucoma (PACG) and to assess the efficacy of medical and surgical treatment in terms of intraocular pressure (IOP) and visual field stabilization. METHODS: Seventy consecutive patients with chronic PACG, whose IOP remained > 21 mmHg despite a patent iridotomy, had their IOP controlled by medications or trabeculectomy performed without antimetabolites. They were followed over a 6-year period. Best corrected visual acuity, IOP (mean of annual diurnal variation readings), cup : disc ratio and visual fields were recorded. A trabeculectomy was performed if the IOP was not adequately controlled on maximal tolerable medical therapy or if there was a progression of the glaucomatous defect. Data from one eye of each patient were analysed; if both eyes met the inclusion criteria, one was randomly selected for the analysis. The baseline parameters were compared with those at the end of 6 years. RESULTS: A total of 46 eyes (65%) were controlled medically throughout the 6-year follow-up period, while 24 eyes (35%) required surgery. The mean IOP was 25.4 +/- 4.9 mmHg at baseline and 15.6 +/- 4.6 mmHg at 6 years follow-up (p < 0.001). Stereoscopic evaluation of the cup : disc ratio did not show a significant change from a mean of 0.6 +/- 0.18 at baseline to a mean of 0.64 +/- 0.2 at 6 years (p = 0.12). Progression of visual field defects was seen in seven eyes (10%), which had statistically larger cup : disc ratios (p = 0.04) and more extensive visual field deficits at the initial assessment (p = 0.04), and which also maintained higher levels of IOP (p = 0.03) over the 6 years of follow-up. CONCLUSIONS: Stable visual fields and good longterm IOP control were seen in 90% of chronic primary angle closure glaucoma eyes on medical/surgical therapy over 6 years.